The proposed studies will expand the Large Scale Digital Cell Analysis System (LSDCAS) as a tool for quantitative analysis to be used in a variety of cell biology model systems. The prototype LSDCAS is an automated microscope system that can perform real-time analysis of many thousands of individual living cells under a range of environmental conditions. Further development of LSDCAS will apply the talents of researchers in the Colleges of Engineering, Medicine, and Liberal Arts at the University of Iowa, and result in biomedical imaging and data modeling/visualization technologies that will be useful in basic biological research. Funds requested will provide a microcomputer cluster and disk storage facility that will allow image segmentation and analysis to be performed in Engineering without impacting data acquisition in Medicine. Recent advances in image segmentation techniques will be incorporated within LSDCAS to provide quantitative analysis in each of three projects chosen to represent types of applications for which LSDCAS is uniquely suited. Cell growth and death as a result of exposure of human tumor cells to Etoposide, a chemotherapeutic agent, will be analyzed using digital images produced by LSDCAS, in order to determine the mode of cell death as a function of drug dose. Intracellular pro-oxidant levels will be measured using vital fluorescent probes in a cell model system designed to study the effects of oxidative stress upon cell killing in a large population of affected cells. Experiments are planned which will analyze changes in cell motility that are known to occur when a particular tumor suppressor gene, maspin, is silenced in metastatic human cancer cells. These experiments will test the hypothesis that alterations in cell-surface substrate attachment lead to these changes in motility. For all three projects, biophysical models based upon nonequilibrium thermodynamics will be developed as an aid in the visualization of the results of these studies, and to predict the outcome of future experiments. Completion of the proposed studies, and the dissemination of the software produced therein, will establish LSDCAS as a tool for use in a variety of other biomedical experiments.